The Molecular Biology Core Laboratory is directed by Dr. Russell. He will be assisted in day-to-day operations by Dr. Jonathan Cohen, who is an expert in high-throughput DMA sequencing. This Core laboratory provides support for acrylamide gel electrophoresis of proteins, DMA sequencing, analysis of gene expression by oligonucleotide microarray hybridization and real-time polymerase chain reactions (PCRs), oligonucleotide procurement, genomic DMA and RNA isolation, and the maintenance and storage of bacterial strains, plasmids, and purified proteins used within this Program Project. Four experienced technicians, Kevin Anderson (100% time), Emily Brown (100%), Jeffrey Cormier (100%), and Scott Clark (25%) will perform the duties associated with this Core. The laboratory facility is located within the Department of Molecular Genetics. For the analysis of proteins, 30 vertical electrophoresis units and 12 multi-outlet power supplies are available. SDS polyacrylamide gels are either prepared (25% of gels) or purchased pre-poured (75% of gels) and run by a single technician (Emily Brown). Following electrophoresis, individual Investigators working on the different Research Projects process the gels for autoradiography, immunoblotting, or protein sequencing. A darkroom that contains a Konica automatic X-ray developer is used to process all autoradiograms and chemilumigrams. The services of the facility are used extensively to assess protein purification, immunoprecipitation from cells, expression of recombinant proteins, and immunoblotting from cultured cells, tissues, and recombinant hosts. These techniques and methods are crucial to our studies on the expression and purification of essentially all genes and proteins with which we work. In addition, the determination of tissue-specific expression patterns of genes under study via immunoblotting in transgenic and knockout mice is heavily dependent on this aspect of the Core.